Abstract

Background

Deletion of the glycosyltransferase bgsA in Enterococcus faecalis leads to loss of diglucosyldiacylglycerol from the cell membrane and accumulation
of its precursor monoglucosyldiacylglycerol, associated with impaired biofilm formation
and reduced virulence in vivo. Here we analyzed the function of a putative glucosyltransferase
EF2890 designated biofilm-associated glycolipid synthesis B (bgsB) immediately downstream of bgsA.

Results

A deletion mutant was constructed by targeted mutagenesis in E. faecalis strain 12030. Analysis of cell membrane extracts revealed a complete loss of glycolipids
from the cell membrane. Cell walls of 12030ΔbgsB contained approximately fourfold more LTA, and 1H-nuclear magnetic resonance (NMR) spectroscopy suggested that the higher content
of cellular LTA was due to increased length of the glycerol-phosphate polymer of LTA.
12030ΔbgsB was not altered in growth, cell morphology, or autolysis. However, attachment to Caco-2
cells was reduced to 50% of wild-type levels, and biofilm formation on polystyrene
was highly impaired. Despite normal resistance to cationic antimicrobial peptides,
complement and antibody-mediated opsonophagocytic killing in vitro, 12030ΔbgsB was cleared more rapidly from the bloodstream of mice than wild-type bacteria. Overall,
the phenotype resembles the respective deletion mutant in the bgsA gene. Our findings suggest that loss of diglucosyldiacylglycerol or the altered structure
of LTA in both mutants account for phenotypic changes observed.

Conclusions

In summary, BgsB is a glucosyltransferase that synthesizes monoglucosyldiacylglycerol.
Its inactivation profoundly affects cell membrane composition and has secondary effects
on LTA biosynthesis. Both cell-membrane amphiphiles are critical for biofilm formation
and virulence of E. faecalis.